The family duo was joined by The Sing-Off champs and Grammy winners Pentatonix. Together, the three artists beautifully blended their voices in the haunting tune.
They all looked good too — wearing a matching mix of silver and black outfits. Cyrus, 24, stood out, in a sheer silver floral and black sun dress with knee-high silver sequence boots.
Parton, meanwhile, was showed off her tiny frame in form-fitting black leather pants with sparkly silver accessories and a long-sleeved black sweater with shoulder cutouts.
This isn’t the first time Cyrus and Pentatonix have covered “Jolene.”
In 2012, the “Party in the U.S.A.” singer covered her godmother’s tune as part of her “Backyard Sessions” series.
Pentatonix, meanwhile, recorded a cover of the track with Parton in September, which was released as a digital single.
“Singing with @PTXofficial was pure magic, and we had lots of fun rearranging the song!” Parton tweeted at the time.
“It was magical singing with YOU!! We will never forget it! <3” they responded.
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Earlier on Tuesday’s episode of The Voice, Parton performed her new song “Circle of Love” — with the help of the show’s Top 10 contestants and country star Jennifer Nettles.
Nettles stars in Parton’s heartwarming holiday sequel and Yuletide adventure, Christmas of Many Colors: Circle of Love. The two-hour made-for-TV movie premieres Wednesday at 9 p.m. ET on NBC.
The Voice airs Mondays and Tuesdays at 8 p.m. ET/PT on NBC.